Expression of chemokine receptors on Th1/Th2 CD4[+] lymphocytes in patients with multiple sclerosis

Th1 cells preferentially express CXCR3, CCR5 and CCR6, while CCR3 and CCR4 are predominantly expressed by Th2 cell subsets. Multiple Sclerosis [MS] is a TH-1 cell-dependant chronic inflammatory disease of the central nervous system, and immunomudolatory cytokines could alter the chemokine expression pattern of these lymphocyte subsets. This study was performed to measure chemokine receptor expression on CD4 T cells for evaluation of Th1/ Th2 dominantly in IFN-beta treated patients. Flowcytometry was used to detect chemokine receptor expression on CD4 T cell population in PBMCs obtained from MS and healthy control groups. Twenty six MS patients participated in this study before and after IFN-beta therapy and the same number of healthy individuals were included. The percentage of lymphocytes was 41.28 +/- 10.30 in the blood of MS group compared with 36.88% +/- 5.51% in the control group [p=0.017]. The CD4[+] CXCR3[+] cells were 18.86% +/- 8.46% in healthy group, 30.78% +/- 9.8% in pre-treated MS patients and 21.06% +/- 9.23% in post-treated group [p<0.001]. The CD4[+]CCR4[+] cell subsets were 27.35% +/- 10.15% in healthy group; 28.17% +/- 8.9% in pre-treated group and 34.20% +/- 8.96% in the post-IFN-beta treatment group. The subset of CD4[+]CCR4[+] was found to be dominant after IFN-beta therapy in comparison with the control group [p<0.001]. CD4[+]CCR5[+] percentage was 1.24% +/- 0.92% in the healthy people, 1.23% =/- 0.71% in the MS patients and 0.76% +/- 0.49% in post-treatment status [p=0.003]. CD4[+]CCR3[+] cell subsets were 0.62% +/- 0.67% in control group, 0.28% +/- 0.26% in the MS patients [p=0.022] and 0.39% +/- 0.54% in IFN-beta treated patients [p=0.334]. An association was found for CXCR3 expression in pre- and post- treatment status [r=0.840, p<0.001] as well as for CCR4[+] expression [r=0.712, p<0.001] in the same groups. The Th1 response was dominant in pre-treatment states, and then the chemokine receptor expression of Th1/Th2 cell subsets could be used for monitoring and the evaluation of the MS disease status

assessment of NK cytotoxicity and CD56+/CD16+ phenotype by flow cytometry in PBL isolated from women with recurrent spontaneous abortion

Human peripheral blood NK cells constitutively express CD56 and CD16 antigens. Peripheral blood NK cells seem to be strongly related with decidual NK cells, and may reflect the decidual NK cell functional status. There are varied reports concerning the relationship between NK cell cytotoxicity in women with recurrent spontaneous abortion. To study NK activity in women with history of RSA by using a non-radioactive cytotoxicity assay. Peripheral blood lymphocytes from RSA and healthy multiparous women were obtained. Lymphocytes were isolated and mixed with K562 NK-sensitive cell line. A non-radioactive method for NK cell cytotoxicity assessment was utilized. Dead K562 cell populations were detected by FACS Calibur flow cytometry, and the data obtained was analysed on cell-Quest software. The proportion of CD16+/CD56+ cells was then calculated. The proportion of NK cells were 9.21% +/- 3.06 and 13.48% +/- 4.09 in healthy women and RSA, respectively. The percentage of cytotoxicity was determined to be 19.3% +/- 3.9 in healthy group and 27.1% +/- 6.5 in RSA group with an effector:target ratio of 50:1. The data shows an increase in PBLs potential for in vitro cytotoxicity assay in RSA individuals. The analyses indicate that there is a weak positive correlation between NK cytotoxicity potential and the percentage of NK cells in PBL population. The present study demonstrates that the percentage of CD56+/CD16+ cells increases in individuals with recurrent spontaneous abortion. We conclude that NK cytotoxicity as well as NK number is partially involved in RSA